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RNA Editing Implicated in Chloroplast-To-Nucleus Communication

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RNA Editing Implicated in Chloroplast-To-Nucleus Communication COMMENTARY RNA editing implicated in chloroplast-to- nucleus communication COMMENTARY Robert M. Larkina,1 Photosynthesis and respiration provide the chemical biogenesis at a stage resembling proplastids, the ex- energy that sustains life on Earth. In eukaryotes, this pression—usually the transcription—of PhANGs is se- essential metabolism is performed by chloroplasts and verely down-regulated. Indeed, when chloroplast mitochondria. Although both of these organelles have biogenesis is blocked in light-grown seedlings, small genomes that are remnants of their endosymbi- PhANGs are expressed at lower levels than are ob- otic origins, most of the proteins in these organelles served in dark-grown seedlings, which contain etio- are encoded by nuclear genes. One consequence of plasts—an intermediate in chloroplast biogenesis that this distribution of genetic material is that many of the accumulates in the absence of light-dependent gene multisubunit protein complexes that are central to expression and chlorophyll biosynthesis. In gun mu- energy metabolism are composed of proteins tants, when chloroplast biogenesis is blocked, PhANG encoded by nuclear genes and either chloroplast expression is significantly up-regulated relative to the genes (photosynthesis) or mitochondrial genes (respi- wild type because of abnormal chloroplast-to-nucleus ration). Thus, the coordination of nuclear and chloro- signaling. The chloroplast-to-nucleus signaling that plast (or mitochondrial) activities requires the depends on the GUN genes is also activated when anterograde flow of information from the nucleus the biogenesis of chloroplasts and etioplasts is not and retrograde signaling back to the nucleus. Al- blocked but their activities are attenuated. When though we know that retrograde signals from chloro- seedlings contain well-functioning chloroplasts, PhANG plasts (i.e., chloroplast signals) influence numerous expression is indistinguishable in gun mutants and wild chloroplastic and extrachloroplastic processes, we still type. Thus, chloroplast dysfunction appears to activate have major gaps in our knowledge of this type of this type of chloroplast-to-nucleus signaling (2). signaling (1, 2). RNA editing refers to processes that gun mutant screens have yielded a large number of change the identities of nucleotides and processes that mutant alleles of genes that encode a plastidic penta- add or delete nucleotides from RNAs. RNA editing has tricopeptide repeat (PPR) protein named GUN1, a blue- been reported in viruses and diverse eukaryotes. In flow- light receptor named cryptochrome 1 (cry1), and ering plants, RNA editing converts cytidines to uridines proteins that contribute to tetrapyrrole metabolism in the RNAs transcribed from the chloroplast and mito- (GUN2, GUN3, GUN4, GUN5, and GUN6), which occurs chondrial genomes (3). In PNAS, Zhao et al. (4) make a in chloroplasts and nonphotosynthetic plastids. In plants, strong case that RNA editing in chloroplasts contributes tetrapyrroles include siroheme, heme, phytochromobi- to a type of chloroplast-to-nucleus signaling defined by lin, and chlorophyll. Increases in the levels of GUN6 the genomes uncoupled (gun)mutantsinArabidopsis. (also known as ferrochelatase 1) activate chloroplast- The gun mutant screen was the first forward ge- to-nucleus signaling, probably by increasing heme netic screen developed that specifically interro- biosynthesis. cry1 can accumulate in either the cytosol gates chloroplast-to-nucleus signaling. The mutant or the nucleus and up-regulates or down-regulates alleles from this screen uncouple the expression PhANG expression depending on factors that influence of photosynthesis-associated nuclear genes (PhANGs) cellular conditions, such as chloroplast function. The from chloroplast function. Chloroplasts develop from specificity of the gun mutant screen indicates that nonphotosynthetic proplastids during the develop- chloroplast dysfunction activates a few specific mech- ment of photosynthetic organs such as cotyledons anisms that regulate gene expression in the nucleus and leaves. Light is a major positive regulator of PhANG and that the gun phenotype is uncommon. A gun expression. When light-grown plants are treated with phenotype refers to the significant up-regulation of inhibitors or harbor mutant alleles that block chloroplast PhANG expression in seedlings containing dysfunctional aKey Laboratory of Horticultural Plant Biology, Ministry of Education, College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070, People’s Republic of China Author contributions: R.M.L. wrote the paper. The author declares no conflict of interest. Published under the PNAS license. See companion article 10.1073/pnas.1820426116. 1Email: [email protected]. www.pnas.org/cgi/doi/10.1073/pnas.1905566116 PNAS Latest Articles | 1of3 Downloaded by guest on September 24, 2021 Well-functioning Dysfunctional photosynthetic cell photosynthetic cell Plastid Plastid YS1 OPT84 GUN1 MORF2 YS1 OPT81 OPT84 MORF2 RNA OPT81 Editing Abnormal RNA Editing Cytosol Cytosol PhANG PhANG Nucleus Nucleus Fig. 1. Proposed interactions between RNA editing and chloroplast signaling. In well-functioning photosynthetic cells (Left), chlorophyll accumulates in the thylakoid membranes (green), and chloroplasts perform photosynthesis. The GUN1 protein does not accumulate and thus the chloroplast-to- nucleus signaling that depends on GUN1 is not active. MORF2 contributes to RNA editing (green arrow) and interacts with OPT81, OPT84, and YS1. Light signaling, tissue-specific signals, and the circadian rhythm (not shown) drive high-level expression of PhANGs (black arrow), which promotes chloroplast function. Blocking chloroplast biogenesis at a stage resembling the proplastid leads to the development of dysfunctional photosynthetic cells (Right) containing dysfunctional chloroplasts that do not accumulate chlorophyll or perform photosynthesis. In these cells, GUN1 accumulates and regulates RNA editing by interacting with MORF2 (red arrow). Abnormal RNA editing activates a chloroplast-to-nucleus signaling mechanism that down-regulates PhANG expression (red T-bar). Similarly, GUN1 accumulates during the early stages of leaf development—before chloroplast biogenesis is completed (11)—and regulates RNA editing and chloroplast-to-nucleus signaling as it does in dysfunctional photosynthetic cells. chloroplasts, presumably because of abnormal chloroplast-to- RNA than wild type (9). Alternatively, based on a variety of ex- nucleus signaling. The GUN genes appear to promote chloroplast periments with overexpressed fusion proteins containing GUN1, function (1, 2). Consistent with this observation, all of the gun mutants such as the cross-linking of an overexpressed fusion protein con- that were isolated from gun mutant screens are more sensitive to the taining GUN1 to nearly 300 different chloroplast proteins, in- inhibitors that are typically used to block chloroplast biogenesis in cluding GUN6, Tadini et al. (10) proposed that GUN1 might these types of experiments (e.g., norflurazon and lincomycin) than wild contribute to plastid signaling by associating with some of these – type (5 7). Norflurazon blocks chloroplast biogenesis by inhibiting the putative GUN1-binding proteins. Although this is an intriguing biosynthesis of carotenoids, which are essential photosynthetic pig- idea, at this stage, the data consistent with this idea are subject ments. Lincomycin blocks chloroplast biogenesis by inhibiting chloro- to interpretation. plast translation (2). Zhao et al. (4) make a strong case that GUN1 contributes to GUN1 is associated with a number of biological functions, such chloroplast-to-nucleus signaling by influencing RNA editing. They as chloroplast biogenesis, the regulation of nuclear gene expres- report that in norflurazon-treated seedlings, RNA editing is al- sion, the circadian rhythm, the accumulation of anthocyanins, abi- tered at 21 sites and that GUN1 affects RNA editing at 11 of these otic stress tolerance, and the development of seedlings and leaves sites in norflurazon-treated seedlings. Importantly, they found that (1, 2). Data from genetic experiments indicate that GUN1 is central GUN1 has only minor effects on RNA editing in untreated seed- to much of this type of chloroplast signaling (1, 2). However, a lings containing well-functioning chloroplasts. Moreover, they particular type of chloroplast signaling that is activated by lincomy- show that GUN1 can interact with MULTIPLE ORGANELLAR cin treatments was recently reported to not depend on GUN1 (8). RNA EDITING FACTOR 2 (MORF2), an essential chloroplast pro- Unfortunately, our knowledge of the biochemical function of tein and a core component of the RNA editosome that edits many GUN1 has been vague. In general, PPR proteins contribute to of the RNAs that are edited in chloroplasts. Although it is not RNA metabolism in chloroplasts and mitochondria. Thus, GUN1 possible to test whether loss-of-function alleles of morf2 influence was suggested to participate in chloroplast-to-nucleus signaling chloroplast-to-nucleus signaling because this mutant is seed- by contributing to RNA metabolism in chloroplasts that affects ling lethal, overexpression of MORF2 induces a gun phenotype. the biosynthesis or transduction of a chloroplast signal (1, 2). Con- Zhao et al. (4) also report that three RNA editing site-specificity sistent with this idea, gun1 mutants accumulate less chloroplast factors named ORGANELLE TRANSCRIPT PROCESSING 81
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